A simple purification scheme yielding crystalline phospholipase C from Bacillus cereus.
A very simple and rather unusual purification scheme for phospholipase C from Bacillus cereus has been worked out. Air is bubbled vigorously through the bacterial culture and the foam collected. Liquified foam is centrifuged, dialyzed and heated at 74 degrees C for 5 min. After centrifugation, affinity chromatography is carried out on lipoprotein-Sepharose. The enzyme is then thermally denatured by exposure to 85 degrees C for 5 min and the precipitated material well washed and then renatured from solution in 4 M guanidinium chloride. The final enzyme preparations are electrophoretically homogeneous and easy to crystallize. The recovery of activity exceeds 80%.
D001409:Bacillus cereus / Q000201:enzymology*; D003460:Crystallization; D006358:Hot Temperature; D010740:Phospholipases / Q000302:isolation & purification*; D011489:Protein Denaturation; D010738:Type C Phospholipases / Q000302:isolation & purification*
D010740:Phospholipases; D010738:Type C Phospholipases